TR201816372T4 - Ethoxyketone compounds for enzyme inhibition. - Google Patents

Abstract

The present disclosure relates to novel compounds and pharmaceutical compositions thereof useful as inhibitors of proteasomes. The compounds provided herein have improved proteasome potential and selectivity and increased aqueous solubility and are useful in the treatment of various disorders or diseases associated with proteasomes.

Description

DESCRIPTION ETOXYKETONE COMPOUNDS FOR ENZYME INHIBITION CAPRAZ REFERENCE TO RELATED APPLICATIONS requests priority against the application.

FIELD OF THE INVENTION The present invention relates to compounds having a backbone of the peptide epoxyketone structure. is relevant. These compounds are useful, for example, in inhibiting proteasomes.

INFRASTRUCTURE The proteasome plays a critical role in mediating the regulated degradation of intracellular proteins. It is a multicatalytic protease complex that plays a role. 268 of the in vivo proteasome complex believed to exist as a proteasome, which in terms of molecular weight about and two 198 consists of regulator particles. The nucleus is hollow and its proteins are inside It provides a closed cavity in which it decays. Each end of the core particle has multiple Polyubiquitinated proteins containing ATPase active domains and ubiquitin-binding domains with a 198 regulator subunit that discriminates and transfers them to the catalytic core is related. An alternative form of the regulatory subunit called the 118 particle essentially related to the nucleus in the same way as 198 particles; 118 foreign proteins may play a role in degradation. Core particle 208 proteasome molecular It is about 700 kDa in weight and consists of 28 subunits organized in four rings. is formed. In yeast and other eukaryotes, 7 0 (subunit each of the two outer rings) and 7 13 subunits form each of the two inner rings. a 0( ring 198 or for the 118 regulator complex, the binding areas are also one for the two 13› inner rings. It acts as a physical barrier. The two inner ß rings contain active proteolytic domains.

The degradation of proteins is centrally formed by the joining of two ß-rings. occurs in the reservoir. Inhibition of 208 proteasomes in vivo 268 It is directly related to the inhibition of the proteasome. There are two forms of proteasomes: in the body ubiquitously expressed by the majority of cells The basic structure is in the proteasome and hematopoietic cells and exposed to inflammatory cytokines. immunoproteasome predominantly expressed in surviving cells. proteasome mediated Protein degradation is highly regulated required for various intracellular processes. it is a done name. The eukaryotic proteasome through the use of different peptide substrates Three main proteolytic activities have been identified for: after large hydrophobic residues cleaved chymotrypsin-like activity (CT-L): trypsin cleaved after essential residues peptidylglutamyl peptide cleaved after similar activity (T-L) and acidic residues hydrolyzing activity (PGPH). The proteasome has long been an attractive target for drug development. It was distinguished long ago and was first clinically used as a therapeutic target in oncology. Peptide boronic acids, β-Lactones and peptide to inhibit proteasome activity Various small molecules have been used, including epoxyketones (Bennett and fully understand the roles of the proteasome at the molecular, cellular, and in vivo levels. from the appropriate specificity and/or potential necessary to investigate and uncover is exempt. For example, the peptide has been found to inhibit other proteases. boronic acids and β-lactones are not specific for the proteasome (Borissenko and Groll, 21, 245-273). This is due to the inhibition of these inhibitors by non-proteasome targets. increases the possibility of exhibiting related off-target activities in vivo. epoxyketones are highly selective as inhibitors of proteasomes. However, this peptide epoxyketones have low aqueous solubility and/or less optimal proteasome inhibitor has potential. Therefore, advanced pharmaceutical and/or biological There is a need to develop new proteasome inhibitors with specific properties.

BRIEF CLICK Provided herein are compounds having a tri-peptide ketone epoxy backbone. Moreover with enhanced water solubility and enhanced proteasome inhibitory properties. compounds are described.

A structure shown here in Formula (I) or an enantiomer thereof, diastereomer, tautomer, pharmaceutically acceptable salt, or solvatina Compounds with or prodrug are described: 2 Formula (i) where R1 is -(CH2)m-R4 where m=0 or 1, R4 is from the group consisting of .1 /( f "47" A "T" NK 'track' x'i) l ,x , w` R5 .' q' ,i Rt` III. Hb", `C !f "Nf.v//'P`x_7/` \ \ ioalkyloxyalkyl is NH2, NHRG, -R7-O(C=O)-R3, -R7-(C=O)X-Ra, -R7-OP03M1M2, R7 XYZ R9 N,, RY-XYZ -Rg- ` N R5 is C1-10alkyl, phenyl, -(C=O)-C1-6alkyl or -(C=O)-phenyl, R7 and Rg are each independently either absent or C1-10alkylene, Rg1 each independently H, hydroxyl, metal or M1 and Mz are each independently H or metal, X is absent or O, Y is absent or -(C=O)-, Z is absent or is and walkoxyalkyl, aryl, C1-10aralkyl, heteroaryl, C1-10heteroaralkyl, heterocyclyl, C1- mheterocycloalkyl is selected from carbocyclyl and 01-10carbocycloalkyl, where R3,R1 and R2 is isobutyl, it is not 4-pyridylmethyl.

Also here to have a configuration of Formula (II) Describes the compound of Formula (I) provided: Formula (ii) wherein R1, R2 and Rs are each as defined above.

R4 may be selected from the group consisting of:

DONLC? HQ› 0& W R1 may be selected from the group consisting of: OH?? [\ /° oûACVx er R2 is Cmoalkyl, can happen. or C1-1oheteroaralkyl C1-1oalkoxyalkyl, aryl, heteroaryl, C1-1oaralkyl R2 may be methyl oxy methyl 4-pyridylmethyl, isobutyl, benzyl or 4-thiazolyl-methyl R3 may be Cmoalkyl, aryl, heteroaryl, Cmoaralkyl or C1-1oheteroaralkyl.

R3 may be isobutyl, 4-pyridylmethyl or benzyl.

In one aspect, the present invention is a tri-peptide capable of inhibiting the catalytic activity of the 208 proteasome. Provides epoxyketones. In certain embodiments, the tri-peptide provided herein to an IC50 inhibition of the CT-L catalytic activity of proteasomes at a concentration of 208. has.

In another aspect, the compounds disclosed herein have improved water solubility. Clear in applications, the tri-peptide epoxyketones provided herein are at least about 0.02, 0.05, 0.1, It has a water solubility of 0.5 or 1 mg/m.

In another aspect, the present invention provides pharmaceutical compositions, an acceptable carrier and among others, for example, cancer, inflammation, neurodegenerative disease (such as Alzheimer's), muscle atrophy disease, chronic contagious related to diseases, fever, inability to use muscles, denervation, nerve damage and immunity useful in the treatment of human conditions, including those described herein contains a therapeutically effective amount of a 208 proteasome inhibitor.

In certain embodiments, pharmaceutical compositions are approximately 10'9g of a compound provided herein. to about 10 g. Appropriate daily dosages per subject are approximately 0.01 mg to about 5 g.

Certain applications require the compounds or pharmaceutical compositions described herein. a parenteral route (eg, subcutaneous, intravenous, intramuscular, intraarterial, intrathecal, intracapsular, intraorbital, intracardiac, intradermal, intraperitoneal, transtrecheal, subcuticular, intraarticular, subcapsular, subarachnoic, intraspinal, intraterna and/or infusion) and a non-parenteral route (for example, oral, enteral, buccal, nasal, intranasal, transmucosal, epidermal, transdermal, dermal, ophthalmic, pulmonary, sublingual, rectal, vaginal or topical) formulated into a dosage form.

Another aspect of the present invention is to treat a condition associated with 208 proteasomes. relates to methods of includes applying the amount.

In another aspect, the present invention is a method of making a peptide epoxyketone. it provides.

Other features and advantages of the present invention are from the following detailed description and will become clear from the prompts.

BRIEF DESCRIPTION OF THE FIGURES FIG. 1, blood collected from BaIb/c mice administered a compound described herein represents the remaining percentage of proteasome activity in the samples. your vehicle The proteasome activity in blood samples collected from mice to which it was administered was 100%. evaluated (control group). The remaining activity is a compound described here. Via intravenous injection (IV) at mg/kg or oral feeding at 30 mg/kg The proteasome in blood samples collected from mice administered via (PO) It was calculated by comparing the activity of the control group with the control group.

FIGURE 2A in mice carrying HT-29 human colorectal adenocarcinoma xenografts It shows the antitumor effect of CX13-103. Statistical analysis 2-way ANOVA obtained using

FIGURE 28, Antitumor effect of CX13-103 in mice carrying RL human Ienfoma xenografts shows its effect. Statistical analysis was obtained using 2-way ANOVA.

DETAIL Compounds and their pharmaceutical salts Generally, herein a structure of Formula (I) or an enantiomer thereof, diastereomer, tautomer, pharmaceutically acceptable salt, or solvatina Compounds with or prodrug are described: Formula (I) where R1 is -(CH2)m-R4 where m=0 or 1, R4 is from the group consisting of C1-1oalkyl is chosen, R5 is each independently: H, hydroxyl, C1-10alkyl, C1-10alkoxyl, C1_ -R7-(C=O)X-R3, -R7-OP03M1M2 here Re is C1oalkyl, phenyl, -(C=O)-C1.3alkyl or -(C=O)-phenyl, R7 and Rg are each independently absent or C1oalkylene, Each of the Rs is independently H, hydroxyl, metal or 01.10alkyl, -C1-10alkylene, -NR10R11 or -OP03M1M2, Rio and Rii are each independently H, C1-oalkyl (eg C1-salkyl) or substituted C1- ioalkyl (for example C 6aikii), M and Mz are each independently H or metal, X is absent or O, Y is absent or -(C=O)-, Z is absent or O and R2 and Rg are each independently CHoalkyl, CHoalkenyl, C1-1ohydroxyalkyl, C1- malkoxyalkyl, aryl, Cmoaralkyl, heteroaryl, C1-6 heteroaralkyl, heterocyclyl, C1- is selected from ioheterocycloalkyl, carbocyclyl and C-1ocarbocycloalkyl, where R3, R1 and R2 is isobutyl, it is not 4-pidylmethyl. In particular, R5 is -R7-O(C=O)-Rg, and R7 and R3 are defined as above. In particular, R7 is absent and R5 is H, C1-10alkyl (eg C1-4alkyl, C1-6alkyl), -C1-10alkylene, - NR10 is selected from R11, where Rio and R11 are defined above. Clear in applications R7 is Cmoalkylene (eg -CH2-, -C2H4-, -C3H7-, etc.), and R8 is H, C1-1oalkyl (eg C1-6alkyl), -Cmoalkylene, -NR10R11 wherein Rm and R11 is defined above.

Alternatively, R5 is -R7-(C=O)X-Rg and R7 and R8 are defined as above.

In certain embodiments, R7 is absent, X is O and R8 is H, metal (eg Na and K), NH4, C1. alkyl (eg C 1-4 alkyl), -C1-10 alkylene and -NRioRii wherein R10 and R11 is defined above. Specifically there is no R7, no X and R8 from -NR10R11 are selected, where R12 and R13 are defined above.

Alternatively, R5 is NH2. NHCOMe, NHCOEt. NHCOC3H7 or NHBo.

|/ N,” cRy-XYZ_ R9_ R7-XYZ-7R9- ,› N ,R7 XYZ R9 or H:C and R7, R9, X, Y and Z are defined as above. Especially most of R7, R9, X, Y and Z. there is one of them. In certain embodiments, none of X, Y, 2, and R7 are present and R9 is Ci- ioalkylene (eg -CH2-, -CzH4-, -C3H7- etc.). Specifically R7, R9 and X have none, Y - is (C=O)- and Z is O. In particular, both R7 and X are absent, R9 is C1-10alkylene (for example - CH2-, -C2H4-, -C3H7- etc.), Y is -(C=O)- and Z is O. Especially R9 and X have neither, R7 is C1-10alkylene (eg -CH2-, -C2H4-, -C3H7-, etc.), Y is -(C=O)- and Z is O. Notably X is absent, R7 and Rg are both independently C1.10alkylene (eg - CH2-, -C2H4-, -CaH7- etc.), Y is -(C=O)- and Z is O. All of R7, R9, X and Z may not be and Y is -(C=O)-. In particular, each of the Rs independently are: H, -CH3, -C2H5, -CaH7, In certain embodiments, each of the R5 is independently: -OH, -OCH3, -OCsz, - In particular, each of the R5 independently are: -CH2OH, -C2H50H, -C3H70H, - CH2OC(=O)CH2N(C2H5)2, Each of R5 is independently: -(C=O)OH, -(C=O)ONa, -(C=O)ONH4, - Alternatively, each of R5 is independently: -CONH2, -CON(CH3)2, .1 ``O i ›_ NH In particular, R1 is selected from the group consisting of: In particular, R2 and R3 are independently C1-10alkyl, C1-10alkoxyalkyl, aryl, heteroaryl, C1-10aralkyl. or C1-10heteroaralkyl.

R2 and R3 are independently -(CH2)nR12, R12 is C1-6alkyl, phenyl, pyridyl, -O-C1-Galkyl, thiazolyl, where n=O, 1, 2, 3, 4, 5. In particular, R2 is methyl-oxy-methyl, 4-pyridylmethyl, isobutyl, benzyl or 4-thiazolyl-methyl. In particular, R 3 is isobutyl, 4-pyridylmethyl or benzyl.

Examples of compounds having a structure of formula (I) are listed below. Includes compounds: 1* 133% where each of the chiral carbons in Formula (I) is independently in the R configuration or 8 configuration As described herein, a compound of Formula (I) has a configuration where R1, R2 and R3 have the definitions provided above R./U\N (S) min Formula (II) According to the present invention, the compounds provided herein are selected from the compounds listed below. a structure and its enantiomers, diastereomers, tautomers, pharmaceutically acceptable contains its salts or solvates CX13-130` cx13403, cx13404.

CX13-105, CX13-107. 9.3' cx13-135 cx13-137. .:one . ;:21/ 2' CX13-603; CX13-606.

CX13-608.

CX13~705 Pharmaceutically acceptable salts are physiologically acceptable and any salt or ester suitable for administration to a targeted patient can happen.

Examples of pharmaceutically acceptable salts include acid addition salts (e.g. hydrochloride, hydrobromide, hydroidodide, nitrate, sulfate, bisulfate, phosphate, acid phosphate, isonicotinate, acetate, lactate, salicylate, pantotheanate, bitartrate, ascorbate, succinate, maleate, genticinate, fumarate, gluconate, glucaronate, saccharate, formate, benzoate, glutamate, methanesulfonate, ethanesulfonate, benzenesulfonate, p-toluenesulfonate and pamoate salts) and base salts (e.g. aluminum, calcium, lithium, magnesium, potassium, sodium, zinc and diethanolamine salts). substituted or unsubstituted saturated hydrocarbon containing x to y carbons in the chain represents groups. possibly substituted but containing at least one double or triple bond, respectively, or denotes unsubstituted saturated aliphatic groups. alkoxy groups include methoxy, ethoxy, propoxy, tert-butoxy and the like. A Thus, an alkyl makes the alkyl an ether or similar to an alkoxy. is a substitute. thereby forming an ether.

As used herein, the term "01.10aralkyl" is a C1-10 alkyl substituted with an aryl group. represents the group.

As used herein, the term "C1-10heteroaralkyl" is a substituted heteroaryl group. It refers to the C1-1oalkyl group.

The term "aryl" as used herein is 5-, 6-, and 7-membered substituted or unsubstituted contains single-ring aromatic groups where each atom of the ring is carbon. 5- to 6-membered structures, more preferably ring structures containing one to four heteroatoms contains rings.

As used herein, the terms "carboxyl" and "carbocyclyl" are non-aromatic denotes a substituted or unsubstituted ring, where each atom of the ring is carbon. to 10-membered ring structures, more preferably ring structures of one to four heteroatoms refers to 3- to 7-membered rings containing is doing. includes compounds. A common method for making a prodrug is desired. selected parts that hydrolyze under physiological conditions to release the molecule. contains. In other embodiments, the prodrug is an enzymatic activity of the host animal. can be translated via

Biological Activity, Selectivity and Solubility The compounds provided herein have biological activity to inhibit the proteasome. has. Inhibition of the 2OS proteasome is by methods known in the art, eg Stein et al., in the U.S. Methods and analyzes as described in application 09/569748 can be determined through Chymotrypsin-like (CT-L) of 208 proteasomes, peptidylglutamyl peptide hydrolyzing (PGPH) and trypsin-like (T-L) activities succiniI-Leu-Leu-VaI-Tyr-AMC, Z-Leu-, respectively, as substrates in the assay buffer. A fluorogenic substrate analysis method using Leu-GIu-AMC and Boc-Leu-Arg-Arg-AMC is measured with. After cleavage from substrates, the free fluorophore 7-Amin0-4- methylcoumarin (AMC) is measured using a fluorometer and the CT- L, PGPH and T-L activity are determined.

The compounds provided herein are useful because they inhibit the catalytic activities of the proteasome. it inhibits.

In certain applications, the compounds provided herein are approximately 500 nM, 250 nM, 100 nM on the CT-L activity of 208 proteasomes when present in shows inhibition (eg, at least 50% inhibition). Here in certain applications CX13-608) shows inhibition of the T-L activity of 208 proteasomes.

In another aspect, the compounds disclosed herein have improved water solubility. Clear in applications, the tri-peptide epoxyketones provided herein are at least about 0.02i 0.05, 0.1, It has a water solubility of 0.5 or 1 mg/m.

In another aspect, the tri-peptide epoxyketones provided herein are used in both advanced water It has both solubility and proteasome inhibition. Here in certain applications the compounds provided have a water solubility of at least about 0.1, 0.5 or 1 mg/mL and at concentrations below about 100 nM, 50 nM, 10 nM or 5 nM When present, it inhibits the CT-L activity of 208 proteasomes.

Uses of the compounds Another aspect of the present invention relates to inhibiting the activities of the proteasome. relates to methods wherein a therapeutically effective amount of a compound provided herein includes its implementation. "therapeutically effective amount" as used herein sufficient to achieve the intended therapeutic effect or activity in the subject receiving the compound. means a quantity of a compound. The effective amount is age, gender, health of the subject. condition, dosage form of the compound, extent of disease in the subject, etc. to various factors such as may change accordingly. A therapeutically effective amount, among others a doctor or physician who prepared the compound taking into account the factors mentioned above can be determined by

Another aspect of the present invention relates to treating a proteasome-associated condition. relates to methods. The method consists of an effective amount of a compound provided herein. includes its implementation. The compounds provided herein include those requested below. conditions or conditions associated with the proteasome, including but not limited to It can be used to treat any of the diseases. Various conditions or diseases are mediated by the catalytic functions of the proteasome. known or believed. Proteasome inhibition cancers, neurotoxic/degenerative diseases, Alzheimer's disease, ischemic conditions, inflammation, immune-related diseases, HIV infection, organ graft rejection, septic shock, antigen inhibition of its presentation, parasitic infection, conditions associated with acidosis, macular degeneration, pulmonary conditions, muscle atrophy diseases, fibrotic diseases, bone and Multiple diseases, including but not limited to hair growth disorders It has been recommended as a prevention and/or treatment. Therefore, the peptide described here proteasome inhibitor compositions such as epoxyketones should treat patients with these conditions. provides a tool for doing so.

Proteasome inhibition has been clinically demonstrated as an antitumor therapeutic strategy.

Thus, the compounds disclosed herein are useful for treating cancers. Treatment Exemplary cancers that can be used include leukemia, lymphoma, myeloma, and hepatocellular carcinoma etc. including carcinomas. Treatment with the compounds described herein Other cancers that can be diagnosed are adrenocortical carcinoma, AIDS-related carcinomas, astrocytoma, bone carcinoma, osteosarcoma, GBM, malignant fibrosis hydrocytoma, melanoma, malignant mesothelioma, Pheochromocytoma, pineoblastoma and supratentorial primitive neuroectodermal tumors, neuroblastoma, uterine sarcoma, bile duct cancer, bladder cancer, breast cancers, gastrointestinal cancers, cervical cancers, colon cancers, rectal cancers, esophageal cancers, eye cancers, ovarian cancers, neck and head cancers, kidney cancers, lip and sickle cavity cancers, lung cancers, nasal cavity and paranasal sinus cancers, penile cancers, transition cell cancers, salivary gland cancers, soft tissue cancers, includes thyroid, parathyroid cancer and vaginal cancers.

Proteasome inhibitors have been associated with NF-kB inhibition.

NF-kB TNF, IL-1, cyclooxygenase, ICAM etc. including inflammatory molecules such as It is a potent transcription factor that mediates the transcription of genes such as

Therefore, the compounds provided herein include without limitation allergy, asthma, a transplanted organ/tissue rejection, auto-immune diseases Iupus, rheumatoid arthritis, psoriasis, multiple inflammatory disorders such as sclerosis and inflammatory bowel diseases or conditions can be beneficial. To a patient in need of the compounds provided herein, this as an optional pharmaceutical composition in an amount effective to treat the conditions applicable.

Proteasome inhibitors reduce the degradation of muscle proteins, therefore muscle It has been found to be beneficial in inhibiting fiber loss and fiber atropy. Therefore The compounds provided herein can treat cachexia and chronic infectious diseases, fever, muscle misalignment. use and denervation, nerve damage, renal impairment associated with acidosis, and hepatitis It can be used to treat muscle atrophy such as In certain applications The compounds provided herein can reduce muscle protein degradation to a patient in need, intracellular slowing protein degradation or degradation of p53 protein in a cell, or as an optional pharmaceutical composition in an amount effective to reduce applicable.

The compounds provided herein also prevent stroke, ischemic damage to the nervous system, neural trauma. (eg percussive brain injury, spinal injury and traumatic injury to the nervous system). multiple sclerosis and other immune-mediated neuropathies (eg Guillain-Barre syndrome) and variants thereof, acute motor axonal neuropathy, acute inflammatory demyelinating polyneurotherapy and Fisher Syndrome), HIV/AIDS, dementia complex, axonomy, diabetic neuropathy, Parkinson's disease, Huntington's disease, multiple sclerosis, bacterial, parasitic, fungal and viral meningitis, encephalitis, vascular dementia, multi- infarct dementia, Lewy body dementia, anterior Iob dementia eg Pick's disease, subcortical dementias (Huntington's or progressive supranuclear palsy), focal cortical atrophy syndromes (such as primary aphasia), metabolic-toxic dementias and infections including but limited to dementias (such as syphilis or chronic meningitis) caused by It can be used to treat non-neurodegenerative diseases and conditions.

The compounds provided herein also include ß-amyloid, the main cause of Alzheimer's disease. upregulation of protein processing associated with extracellular accumulation of protein (ß-AP) may be beneficial in Therefore, the compounds provided herein Alzheimer's disease, for example, by reducing the rate of ß-AP processing, the rate of ß-AP plaque formation. by reducing the rate of formation of ß-AP and clinical improvement of Alzheimer's disease. It is useful to treat by reducing the symptoms.

Proteasome inhibition has also been associated with reducing fibrosis. So here compounds provided diabetic neuropathy, glomerulosclerosis, IgA nephropathy, cirrhosis, biliary atresia, congestive cup disease, scleroderma, radiation-induced fibrosis, lung for the treatment of fibrosis-related conditions such as fibrosis and cardiac fibrosis can be used.

Pharmaceutical Compositions and Application In another aspect, the present invention provides a pharmaceutical composition, wherein the a compound and a pharmaceutically acceptable carrier.

The term "pharmaceutically acceptable carrier" as used herein a provided compound from a location, body fluid, tissue (internal or external) or from part of the body to another location, body fluid, tissue, organ or body a liquid or solid filler part of which is included in the carriage or transport, pharmaceutically acceptable as diluent, excipient, solvent or encapsulating material means an acceptable material, composition or tool. pharmaceutically acceptable carriers are excessive toxicity or adverse effects to an animal's tissues vehicles, diluents, excipients or other materials can be. Examples of pharmaceutically acceptable carriers are sugars, starch, celluloses, malt, tragacanth, gelatin, Ringer's solution, alginic acid, isotonic saline, buffering agents, etc. contains.

Each carrier may be combined with other components such as the composition of the formulation provided here. compatible and with the tissue or organ of a biological subject, excessive toxicity, irritation, allergic contact without response, immunogenicity, or other problems or complications being proportionate to a reasonable benefit/risk ratio, suitable for use in Relating to materials that can function as pharmaceutically acceptable carriers some examples include: (1) sugars such as lactose, glucose and sucrose; (2) corn starches such as starch and potato starch; (3) sodium carboxymethyl cellulose, ethyl cellulose and its derivatives, such as cellulose and cellulose acetate; (4) pulverized tragacanth; (5) malt; (6) gelatin; (7) talc; (8) such as cocoa butter and auxiliary candles excipients; (9) peanut oil, linseed oil, safflower oil, sesame oil, olive oil, oils such as corn oil and soybean oil; (10) glycols such as propylene glycol; (11th) polyols such as glycerin, sorbitol, mannitol and polyethylene glycol; (12) ethyl oleate and ethyl Iaurate esters such as; (13) agar; (14) such as magnesium hydroxide and aluminum hydroxide buffering agents; (15) alginic acid; (16) pyrogen-free water; (17) isotonic saline; (18) Ringer solution; (19) alcohol such as ethyl alcohol and propane alcohol; (20) phosphate buffer solutions and (21) used in pharmaceutical formulations such as acetone, other non-toxic compatible substances.

Pharmaceutical compositions as pharmaceuticals necessary for physiological conditions such as pH adjustment. acceptable excipients and buffering agents, toxicity modulation agents and the like, such as sodium acetate, sodium chloride, potassium chloride, calcium chloride and sodium lactati and the like.

Pharmaceutical composition solid dosage form (for example, tablets, capsules, powders, granules and etc.) and liquid dosage forms (eg aqueous solution, emulsion, elixir, syrup, etc.) in any suitable dosage form. pharmaceutical compositions Methods of preparation are well known in the art and are for example Remington: The Science and Practice of Pharmacy (Gennaro ed. 20th edition, Williams & Wilkins PA, USA) (2000).

In certain embodiments, the pharmaceutical composition is approximately 10'9 g to about 10'9 g of a compound provided herein. contains about 10 g (for example, about 0.01 mg to about 10 g, about 0.1 about 500 mg or about 10 mg to about 300 mg). Daily available per subject dosages can be from about 0.01 mg to about 5 g.

Certain applications require the compounds or pharmaceutical compositions described herein. a parenteral route (eg, subcutaneous, intravenous, intramuscular, intraarterial, intrathecal, intracapsular, intraorbital, intracardiac, intradermal, intraperitoneal, transtrecheal, subcuticular, intraarticular, subcapsular, subarachnoic, intraspinal, intraterna and/or infusion) and a non-parenteral route (for example, oral, enteral, buccal, nasal, intranasal, transmucosal, epidermal, transdermal, dermal, ophthalmic, pulmonary, sublingual, rectal, vaginal or topical) formulated into a dosage form. Specific examples described here compounds or pharmaceutical compositions that are suitable for delivery to a balance in need It is formulated in dosage form.

Suitable dosage forms are without limitation injectable emulsions, solutions and formulations for parenteral use such as suspensions, tablets, capsules, Formulations for oral use such as pills, dragees, powders and granules, Sprays, such as ointments, pastes, creams, Lotions, gels, solutions, patches, and inhalants formulations for topical or transdermal use and intrarectal formulations such as suppositories. or formulations for intravaginal use. These formulations compound in appropriate conditions with appropriate excipients, Remington: The Science and Practice of By Pharmacy (Gennaro ed. 20th edition, Williams & Wilkins PA, USA) (2000) by joining according to methods known in the art as described can be prepared.

Pharmaceutical compositions are a balance of oral, intravenous, intranasal, topical, intramuscular, by any suitable route, such as via the intradermal, transdermal or subcutaneous routes. can be applied in the form of application route.

In certain applications, the compounds or pharmaceutical compositions provided herein may require such that combining or even synergistic pharmaceutical effects can be achieved in the subject It can be administered with a second agent. For example, the compounds provided herein and The second active agent may be administered as a single pharmaceutical composition or co-administered as separate compositions. It can be applied sequentially or in separate compositions. Cancer The second active agent that can be co-administered with the compounds of the present invention for the treatment of Includes, but is not limited to: 5-fluorouracil, doxorubicin, daunorubicin, tamoxifen, Iuprolide, goserelin, flutamide, niluthimide, finasteride, dexamethasone, lenalidomide aminoglutethimide, amsacrine, anastrozole, asparaginase, beg, bicalutamide, bleomycin, buserelin, busulfan, campotecin, cabecitabine, carboplatin, carmustine, chlorambucil, cisplatin, cladribine, clodronate, colchicine, cyclophosphamide, temozolomide, cyproterone, cytarabine, dacarbazine, dactinomycin, daunorubicin, dienestrol, diethylstilbestrol, docetaxel, doxorubicin adriamycin, epirubicin, estradiol, estramustine, etoposide, exemestane, filgrastim, fludarabine, fludrocortisone, fluorouracil, fluoxymesterone, flutamide, gemcitabine, genistein, goserelin, tamoxifen, teniposide, testosterone, titanocene dichloride, topotecan, trastuzumab, tretinoin, vinblastine, hydroxyurea, idarubicin, ifosfamide, imatinib, interferon, irinotecan, ironotecan, Ietrozole, Iokovorin, pentostatin, plisamycin, porfimer, procarbazine, raltitrexed, rituximab, streptozocin, suramin, Iöprolid, mercaptopurine, mesna, methotrexate, mitomycin, mitotane, mitoxantrone, nilutamide, nocodazole, octreotide, oxaliplatin, paclitaxel, pamidronate, thioguanine, thiotepa, titanocene dichloride, topotecan, trastuzumab, tretinoin, vinblastine, vincristine, vindesine, and vinorelbine.

In certain embodiments, a compound of the invention is a chemical treatment for cancer. can be applied in conjunction with other methods. Available in certain apps A compound of the invention may be administered in conjunction with radiation therapy. Clear A compound of the invention available in applications is surgical, thermoablation, focused ultrasound therapy, chiropractic, or in combination with any of these applicable.

In certain embodiments, a compound of the invention is amcinonide, beclomethasone, betamethasone, budesonide, chloroprednisone, clobetasol, clocortolone, cloprednol, corticosterone, cortisone, sortiazole, desonide, desoxymethasone. dexamethasone, diflorason, diflucortolone, difuprednate, enoxolone, fluazacort, flumethasone, flunisolide, fluchloronide, fluocinolone acetonide, fluocinonide, fluocorinbutyl, fluocortolone, fluorometholone, fluperolone acetate, fluprednidene acetate, fluprednisolone, flurandrenolide, fluticasone propionate, formocortal, halobetasol propionate, halcinonide, halomethasone, hydrocortisone, Ioteprednol etabonate, mazipredon, medrizon, meprednisone, methylprednisolone, mometasone furoate, containing paramethasone, prednisolone, dexamethasone and 25-diethylaminoacetate but it is administered in conjunction with a steroid that is not limited to these.

A compound of the invention present in certain applications is verapamil, rapamyscin, mycophenylate MDR such as mofetil, thalidomide, cyclophosamide, cyclosporine, and monoclonal antibodies It is administered with an immunotherapeutic agent, including modulators. EXAMPLES The following are the tri-peptide epoxy ketones described herein, their synthesis, proteasome activity inhibitor property, anti-tumor activity property and their are examples of its solubility in water.

The synthesis of the compounds provided herein is carried out in the synthetic schemes in the examples below. is shown. These diagrams are illustrative only and include the compounds provided here. restricting other possible methods that can be used to prepare is not intended. In addition, the steps in the skies are for better illustration and suitable can be changed in sequence. Applications of the compounds in the examples, research purposes and has been synthesized abroad for potential supply to regulatory bodies.

1H-NMR to obtain information on the structures of compounds for general test methods spectra are used. Liquid chromatography mass spectrometry (LC-MS) positive A quadrupole mass spectrometer operating in the electrospray ionization mode (ESI) taken using the system. High performance liquid chromatography with purity UV detection (HPLC). Preparer high when needed performance liquid chromatography (preparative HPLC) was used for separation purification.

Synthesis of compound CX13-103 Sema 1. Synthesis of Compound CX13-103 0034 cium r 0 4:.` --4 .'\ _ _ , ~ : c i &au; C? I 1)*** "r" _”- cm: 6 cwzs :103.7 cms-a sac-un ' .-. "* l`- [ V ›~ gross“ to x ,1404 ^ ' `v N' - inner 7' "UC" t. .77. 3;" Hrii r...=›\ i-,mF '- (310340 CIUU 11 ii_-' ". 4 r" x_ g# ll : _ L/iun_ \_ -g _ii .: /L CM HM_ ciiu. ;mi .7 r., ` (310343) iim- ii" "CRANE" \_ &JANJL ,dice .'N'x_^ V 6x1:) 103 4-" "x Synthesis of compound C103-2 SOCl2 (5 mL) was added dropwise to MeOH (at 0 °C. Then 0103-1 (10 g, 42 mmol) was added, the mixture was heated under reflux overnight. Room After cooling to temperature, the mixture was concentrated and the crude product C103-2 (11 g, quanti.). MS (ESI) m/z: 181 [M+H]+.

Synthesis of compound C1 O3-6 CCI4 ( NBS to solution (11.5 9, 65 mmol) and AIBN (820 mg, 5 mmol) were added. The reaction mixture was refluxed overnight. heated under flow. After cooling to room temperature, the reaction mixture was dissolved in water. spilled; The 3qu layer was extracted twice with DCM. combined layer washed with saturated NaHCOa, dried over Na2804 and concentrated in vacuo purified by chromatography. MS (ESI) m/z: 221 [M+H]*.

Synthesis of compound C103-7 To a cooled (0 °C) solution of morphine (4.8 9. 55 mmol) in THF (20 mL) The reaction mixture was stirred at 0 °C for 30 minutes. reaction mixture water The aqueous layer was extracted twice with EtOAc. combined layer was washed with brine, dried over N82804 and concentrated in vacuo. purified by chromatography. MS (ESI) m/z: 227 [M+H]+.

Synthesis of C103-8 2 N LiOH (13 mL, 26 mmol) was added dropwise to the solution. Occurring The mixture was stirred at room temperature for 1 hour. pH of aqueous hydrochloride was adjusted to 5 by addition and the mixture was concentrated in vacuo. Residue Purified by chromatography to give C103-8 (2.65 g, yield: 71%). MS (ESI) m/z: 213 [M+H]+.

Synthesis of C103-10 HATU ( added. Mixture 30 minutes The resulting solution was stored under nitrogen at room temperature for 1 hour. mixed. The reaction mixture was poured into water and extracted twice with EtOAc. has been made. The combined layers were dried over Na2804, filtered and ground. Purified by preparative HPLC. MS (ESI) m/z: 442 [M+H]*.

Synthesis of C103-11 TFA (10) into a solution of Compound C. mL) was added slowly at 0 °C. Thereafter, the reaction mixture is It was provided to mix at room temperature throughout. Concentration raw under vacuum yielded the product, which was used for the next step without further purification. MS (ESI) m/z: 342 [M+H]*.

Synthesis of C103-12 HATU ( added. Mixture 30 minutes was stirred at 0 °C throughout. 0103-11 has been added. The resulting solution 1 hour stirred under nitrogen at room temperature throughout. reaction mixture into water poured and extracted twice with EtOAc. Combined layers It was dried over Na2804, filtered and concentrated in vacuo. residual oil (ESI) m/z: 536 [M+H]*. Synthesis of 0103-13 mmol) solution, 2 N LiOH ( was added. The resulting mixture was 1 stirred at room temperature for one hour. Addition of pH aqueous hydrochloride set to 5 via and concentrated in vacuo. Residue C103-13 (3.4 9, yield: 66%) by chromatography. MS (ESI) m/z.' 522 Synthesis of CX13-103 HATU ( added. Mixture 30 minutes PCT/US added. The resulting solution 1 stirred under nitrogen at room temperature for one hour. reaction mixture water poured into it and extracted twice with EtOAc. Combined layers It was dried over Na2804, filtered and concentrated in vacuo. residual oil has been purified. 1H-NMR (, 7.20- m/z: 675 [M+H]+; purity >95%”.

Synthesis of compound CX13-104 Synthesis of intermediate C104-7l Scheme 2: preparation of intermediate C104-7 / C BOC_}O û / 0/ / o ßMgBr c104-1 0104-2 /Ns O vanadil I N" /4O NaBH; › / OH acetiiasetcnat / OH DMF' CeCL; Wi-Buoyia C1040 C1044 CW” Synthesis of C104-1tin solution and 200 ml of THF were added. 20 ml di-t-butyl dicarbonate while stirring was added dropwise and the mixture was stirred for 2 hours. Occurring 300 ml of water was added to the reaction mixture and the resulting mixture was ethyl acetate. (2x200 ml). To adjust the pH of the mixture to the aqueous layer to about 4 a 5% aqueous solution of potassium hydrogen sulfate was added and then the mixture was n- extracted with butanol (4x200 ml). Organic layer under reduced pressure is concentrated. The solid was collected by filtration and as a white solid 8.0 9 Synthesis of C104-02 It dissolves in room temperature and reaches 4 °C. has been cooled. N,O-dimethylhydroxylamine hydrochloride (4.4 9, 45 mmol) was added, followed by HBTU (17 g, 45 mmol) followed by HOBt ( ).

The reaction was allowed to warm up slowly to room temperature and was mixed throughout, then diluted with EtOAc (300 ml) and successively washed with saturated sodium chloride (2x100ml). with organic layer Na2804 dried, filtered and concentrated under reduced pressure. Colon purification by chromatography (DCM (%1TEA)) as yellow oil (6.2 9, 66%) Synthesis of C104-3 The compound dissolved in C and cooled to 0 °C. refrigerated (ice plus salt water). Isopropenyl magnesium bromide (0.5 M in THF; 200 ml; 100 mmol) was added slowly over 20 minutes. Reaction 4 hours was stirred at 0 °C throughout, then a saturated aqueous ammonium chloride at 0 °C. poured into it slowly. THF was removed under reduced pressure and the mixture was extracted with EtOAo (3x100 ml). Combined organic layers saturated sodium bicarbonate was washed with water and saturated aqueous sodium chloride. Organic layer dried with Na2804, filtered and concentrated under reduced pressure. has been made. Purification by column chromatography (DCM (%1TEA)) yellow oil (5.0 9, Synthesis of C104-4 has been cooled. Cerium (III) chloride heptahydrate (8.2 9, 22 mmol) was added, followed by followed by the addition of sodium borohydride (0.85 9, 22 mmol). reaction mix 3 stirred at 0 °C for one hour. The mixture is a saturated aqueous ammonium chloride at 0 °C. (200 ml) and extracted with EtOAc (4x100 ml).

The combined organic layers are then saturated with sodium bicarbonate, water and washed with sea water. The organic layer was dried with Na2804, filtered and concentrated under reduced pressure. Column chromatography (DCM (%1TEA)) Purification via yellow oil ( m/z.' 293.4 [M+H]*.

C104-5lin synthesis The compound dissolved in C and cooled to 0 °C. has been cooled. Vanadyl acetylacetonate (0.12 9, 0.45 mmol) and then tert-Butyl hydroxyperoxide (5.0-6.0 M in decane; 12 ml, 60 mmol) was added. Your reaction allowed to warm up slowly to room temperature overnight.

The mixtures were then washed with saturated sodium bicarbonate, water and seawater.

The organic layer was dried with N82$O4, filtered and any under reduced pressure to give C104-5li used without purification is concentrated. MS (ESI) m/z.' 309.4 [M+H]*.

Synthesis of C104-6 The compound dissolved in C and cooled to 0 °C. has been cooled.

Dess-Martin periodinane (13 g, 30 mmol) was added. room temperature of the reaction and After 24 hours, it was allowed to warm up slowly. My wife later 4 Cooled to °C and more Dess-Martin periodinane (6.4 9. 15 mmol) has been added. Allow the solution to warm slowly to room temperature. has been provided. After 3 hours the mixture was filtered through Celite, the filtrate was then with saturated sodium hyposulfite, saturated sodium bicarbonate, water and seawater has been washed. Organic layer dried with N82804, filtered and reduced pressure concentrated below. Via column chromatography (DCM (%2MeOH)) ivis (ES1) m/z: 307.4 [M+H]*.

Synthesis of C104-7 The compound dissolved in C and cooled to 0 °C. has been cooled. TFA (50 ml) was added. Allow the reaction to cool to room temperature slowly. allowed to warm up. To give C104-7 as the mixture TFA salt concentrated under reduced pressure. MS (ESI) m/z: 207.4 [M+H]*.

Scheme 3: Preparation of CX13-104 CI `1.9' Ü . w:) 4( *i H A cioss "BTU 7% 3.2 1.1”*U' / 5;( th” k r ) D I - en. ' "L-N) U 1/ :i J E FM' Synthesis of Ci 04-8 toluenesulfonic acid monohydrate (9.5 to 9.55 mmol) and toluene (a mixture of Dean- Heated under reflux for 16 hours with a Stark trap. Benzene solution 5% 5qu NaHCO3, washed with brine, dried over MgSCO4, filtered and evaporated to dryness under reduced pressure. Residue colorless needles as C m/z: Synthesis of C104-9 13 mmol) was added to the mixture with DlEA (3 mL) via an addition funnel. Reaction It was kept at the same temperature for 16 hours and then EtOAc (20 mL) and brine. (20 mL). The layers were separated and the aqueous layer with EtOAC ( is extracted. Organic layers combined and on N32804 has dried. Removed by solid filtration and volatiles at reduced pressure removed below to give C104-9 (2.4 9, 61% yield) as a white solid. Purified by silica gel chromatography (PE:EA = 5:1). MS (ES1) m/z: +.

Synthesis of C104-10 mixed below. The mixture is filtered and without purification in the next step. used *.

Synthesis of C104-11 (an addition funnel added via. The reaction was kept at room temperature for 16 hours and It was then diluted with EtOAc (20 mL) and brine (20 mL). aqueous layer EtOAc Extracted with (. Organic layers combined and Na2804 dried on. Removed by solid filtration and volatiles was purified by silica gel chromatography (PE:EA = 5:1) to give MS Synthesis of C104-12 mixed under the atmosphere, filtered and without purification in the next step. used C+.

Synthesis of CX13-104 (the mixture was kept at -5 °C and DIEA was added (1 mL). The reaction was kept at -5 °C for 0.5 hours and then It was diluted with EtOAc (10 mL) and brine (10 mL). The layers are separated and the aqueous layer Extracted with EtOAc (. Organic layers combined and Na2804 dried on. Removed by solid filtration and volatiles removed under reduced pressure. The crude product is compound CX13-104 (50 mg, through purified. 1H-NMR ( Synthesis of compound CX13-105 Scheme 4: Preparing the CX13-105 / HN»O\ / VMQBF EDCI. HOBt N / OH CbZHN ` THF CtiZHN \ CeCIS. MGOH CIVHN DIEA DMF O` 0 û V<){;rii:a<:`i2 Dcss-Marlin periodinan \ Ch?“ N \ DCM l-BUOEH CblHN o DC“ Cb7HN 0 C105-5 C105-G % pass H2 is 5 atm) (3105-7 c1os-7 0 &N 0 n HATU, DIEA, DMF I CX13i105 Synthesis of C105-2 has been added. The mixture was stirred at 0 °C for 30 minutes, then N, O- dimethyl-hydroxylamine hydrochloride (3.2 9, 33.2 mmol) was added. Occurring The solution was stirred for 1 hour at room temperature under nitrogen. Reaction The mixture was poured into water and extracted twice with EtOAc. combined The layers were dried over Na2804, filtered and concentrated in vacuo.

A silica gel column to give the title compound of residual oil C105-2 (8.0 9, yield: 72%?) through purified. MS (ESI) m/z.' 343 [M+H]*.

Synthesis of C105-3 isopropenyl magnesium bromide (added at 0 °C.

The resulting mixture was stirred at 0 °C for 3 hours. reaction mix saturated NH4Cl was poured slowly into the solution. pH level of the solution adjusted to 1.5 using concentrated HCl. The mixture is extracted with EtOAc has been made. The combined layers were dried over Na2804, filtered and ground. vacuo is concentrated. Residual oil C105-3 (5.8 9, yield: 77%) cap compound was purified by a silica gel column to give

Synthesis of C105-4 has been added. The resulting solution was under nitrogen at 0 °C for 3 hours. mixed. Water (5 mL) was added to quench the reaction. In vacuo all After removal of volatiles, the residue was extracted with EtOAc. Combined The filtered layers were dried over Na2804, filtered and concentrated in vacuo. purified by column. MS (ESI) m/z: 326 [M+H]*.

Synthesis of C105-5i mmol) was added. The resulting mixture was kept at room temperature for 2 hours. mixed. Water (5 mL) was added to quench the reaction. Mixed Celite filtered through. The aqueous layer was extracted with DCM. combined sheets were washed with aqueous sodium bisulfite solution and brine, over Na2804 dried, filtered and concentrated in vacuo. Residual oil purification used for the next step without MS (ESI) m/z: 342 [M+H]*.

Synthesis of C105-6 Dess-Martin to a refrigerated (0 °C) solution of 0105-5 in DMF (20 mL). periodinane (13 9, 32.5 mmol) was added. The resulting mixture remains in the room for 1 hour. mixed at temp. The resulting mixture is filtered through celite, The filter cake was washed twice with DCM. Combined layers aqueous sodium bisulfite solution, washed with saturated sodium bicarbonate brine, on Na2804 dried and concentrated in vacuo. Residual oil 0105-6 (2 g, yield: 29%) Purified by silica gel column to give the title compound. MS (ESI) m/z: 340 [M+H]+.

Synthesis of C105-7 Palladium on charcoal (400 mg, 40% by weight) was added to the solution.

The suspension was stirred at 0 °C for 6 hours under a hydrogen atmosphere (3.5 atm). and then the reaction mixture was filtered through celite. Filtrate next step in vacuo to provide the desired crude product that is used without further purification for is concentrated. MS (ESI) m/z: 206 [M+H]*.

Synthesis of CX13-105 mg is added. Mixed 30 minutes was stirred at 0 °C throughout. A solution of C-105-7 in DCM (2 mL) was added.

The resulting solution was stored under nitrogen at room temperature for 1 hour. mixed. The reaction mixture was poured into water and extracted twice with EtOAC. has been made. The combined layers were dried over Na2804, filtered and ground. vacuo is concentrated. To give residual oil CX13-105 (70 mg, yield: 16%) Purified by preparative HPLC. 1H-NMR (, 4.55 1H), m/z: 674 Synthesis of compound CX13-107 Scheme 5: Preparing the CX13-107 m› 1› think.. ciii i. an' '7:' 'w '- x- ~- N - "v * M ru.. /., h .Me -1 Pretty! " i** TFA ::cu ii Ha'u '_ii›_.r«. uii1p~ C 10 7 6 (3107-7 44 H_ I ii..

C x 1 34 07 Synthesis of C107-3 has been added. The mixture was stirred at 0 °C for 30 minutes, then C107-2 (6.2 9, 34.1 mmol) was added. The resulting solution was stored at room temperature for 1 hour. stirred under nitrogen. The reaction mixture was poured into water and mixed with EtOAc. once extracted. The combined layers were dried over Na2804, filtered and concentrated in vacuo. Residual oil C1O7-33 (13.5 9, yield: (ESI) m/z.' 351 [M+H-55]*.

Synthesis of Ct 07-4 TFA (20 mL) was added. The reaction mixture is at room temperature for 1 hour. mixed. Concentration under vacuum gave the crude product, which is another used for the next step without purification. MS (ESI) m/z: 307 [M+H]+ Synthesis of C107-6 HATU ( added. Mixture 30 minutes was stirred at 0 °C throughout. Later, C107-4 was added. Occurring The solution was stirred for 1 hour at room temperature under nitrogen. Reaction The mixture was poured into water and extracted twice with EtOAc. combined The layers were dried over Na2804, filtered and concentrated in vacuo.

Flash on silica gel to give residual oil C107-6 (4.5 9, yield: 88%). purified by chromatography. MS (ESI) m/z: 434 [M+H]*.

Synthesis of C107-7 mmol) solution was added 2 N LiOH (10 mL, 20 mmol). The resulting mix 1 stirred at room temperature for one hour. Addition of pH aqueous hydrochloride via is set to 5. The resulting solid was washed with collected water and It was dried under vacuum to give compound (3.8 9, yield: 90%).

Synthesis of CX13-107 mmol), HATU ( C-105-7 into solution has been added. The resulting solution was stored under nitrogen at room temperature for 1 hour. mixed. The reaction mixture was poured into water and extracted twice with EtOAc. has been made. The combined layers were dried over Na2804, filtered and ground. vacuo is concentrated. to give the residual oil CX13-107 (70 g, yield: 16%) Purified by preparative HPLC. 1H-NiviR (, 4.36-4.31 (m, m/z: 607 [M+H]*. purity is >95%.

Synthesis of compound CX13-130 Scheme 6: Preparing the CX13-1301 HIHwIIFm ..i/s::. one/ \,-, A› ""v'fîçgx .__ ,:;i _end-ç?, H " V \ iirz'ii l'Jll'A 1 T F's'Ü (1st; 11%..- .I ti'- J\/I"J}i _ ' . Bac"`~` 3" "v/"3nd" › E'SCNN --..- CH DV: i', BÜCHN H "î/`i`iig' .'_A ( F1 ,A ,N, KI.” /"'H / "u/K" V" \.,i` Hijiaz :IILA CMS-130 N Synthesis of C130-2 Compound C-benzyl 2- amine and DIEA (4 mL) was added at room temperature. The reaction is at the same temperature for 16 hours. was retained and then diluted with EtOAc (80 mL) and brine (40 mL).

The layers were separated and the aqueous layer was extracted with EtOAc (. Organic The layers were combined and dried over N82804. through solid filtration removed and the volatiles removed under reduced pressure to form a white solid. through purified. MS (ESI) m/z.' *.

Synthesis of C130-3 was added and the mixture was stirred for 16 hours at 50 °C under an H2 atmosphere.

The mixture was filtered and used without purification in the next step, 0130-3 (1100 mg, Synthesis of C130-4 The compound C-2-amino-4- mmol), HOBt ( added. Reaction 16 hours was stirred at room temperature throughout, and then EtOAc (60 mL) and brine (20 mL) was diluted with. The layers were separated and the aqueous layer was extracted with EtOAc ( has been made. The organic layers were combined and dried over Na2804. Thick removed by filtration and volatiles under reduced pressure to give compound C130-4 (510 mg, 34% yield) as a white solid which was removed silica gel chromatography (PE:EA= m/z.' *.

Synthesis of C130-5 Compound C 0 It was added at °C. The reaction was held at 0 °C for 2 hours and the volatiles turned white. removed below. MS (ESI) m/z: *.

Synthesis of CX13-130 Acetic acid to the mixture in Compound C (14 mg, and DlEA (0.2 mL) was added at room temperature. The reaction is at room temperature for 16 hours. was retained and then diluted with EtOAc (40 mL) and brine (10 mL).

The layers were separated and the aqueous layer was extracted with EtOAc (. Organic The layers were combined and dried over Na2804. through solid filtration removed and the volatiles removed under reduced pressure to form a white solid. silica gel chromatography (DCMzMeOH =) to give CX13-130 (25 mg, 21%) as 4011) and purified by preparative HPLC. EXAMPLE Synthesis of compound CX13-133 Scheme 7: Preparation of CX13-133 / [JA 't ü\/› \/\/n`*~ç."\ 't Ü / WC'H CX13-133 Synthesis of C133-2 were mixed at 50 °C throughout. Removed by solid filtration and volatiles C133-2 (1.0 9, 81%) removed under reduced pressure and as a colorless oil via silica gel chromatography (PE:EA=4:1) to give compound has been purified. MS (ESI) m/z.' *.

Synthesis of C133-3 mixed at room temperature. Then the mixture was acidified to pH = 5, concentrated to give C133-3 (750 mg, 84% yield) as a white solid. purified by chromatography. MS (ESI) m/z: *.

Synthesis of CX13-133 To its mixture in Compound C 133-3 (54 mg, added at room temperature. The reaction was kept at room temperature for 16 hours. and then diluted with EtOAC (40 mL) and brine (10 mL). The layers are separated and the aqueous layer was extracted with EtOAc (. Organic layers combined and dried over Na2804. removed by solid filtration and The volatiles were removed under reduced pressure and CX13-133 as a white solid. silica gel chromatography (DCMzMeOH = 40:1) and preparative to give (55 mg, 41%) Purified by HPLC. 1H NMR ( Synthesis of compound CX13-135 Scheme 8: Preparing the CX13-135 DMF,, cx13135 |\:N 5- to its mixture in Compound C methylisoxazoI-S-carboxylic acid (40 mg, , HOBI (39 mg added at room temperature).

The reaction was kept at room temperature for 16 hours and then EtOAc (40 mL) and diluted with brine (10 mL). The layers were separated and the aqueous layer was extracted with EtOAc (. The organic layers were combined and over Na2804). has dried. Removed by solid filtration and volatiles at reduced pressure removed below to give CX13-135 (18 mg, 16%) as a white solid. via silica gel chromatography (DCM:MeOH = 4021) and preparative HPLC has been purified. 1H NMR (, Synthesis of compound CX13-137 Scheme 9: Preparation of CX13-137 -1 0 "r" &"33 J\ 1.' Q 1305 Ltd/f* EMF " `J Mixture of 5-methylthiazol-2- in Compound C carboxylic acid (41 mg, , HOBt (39 mg, 0.29) mmol) and DIEA ( added at room temperature. The reaction was carried out at room temperature for 16 hours). temperature and then with EtOAc (40 mL) and brine (10 mL). it is diluted. The layers were separated and the aqueous layer was extracted with EtOAc ( has been made.

The organic layers were combined and dried over N82804. solid filtration and the volatiles were removed under reduced pressure and by chromatography to give CX13-137 (36 mg, 31%) as a white solid has been purified. 1H NMR (, 7.41 (dd, J = 29.1), Synthesis of compound CX13-600 Scheme 10: Preparing the CX13-600 W” 1“ .W «721 ;r s w. yas/k" ”x/kûa ._ fr“ .. M` ..

CX13-600 Synthesis of CöOO-Z C600-1 (1.38 9, added at room temperature.

The reaction was kept at room temperature for 16 hours and then EtOAc (80 mL) and diluted with brine (40 mL). The layers were separated and the aqueous layer was extracted with EtOAc (. The organic layers were combined and over N32804. has dried. Removed by solid filtration and volatiles at reduced pressure compound C600-2 (1.8 9, 69% yield) as a white solid, which was removed under silica gel chromatography (PE:EA= CoOO-Sun synthesis mixed. The mixture was filtered and used without purification in the next step. 3 ( m/z.' *.

Synthesis of the CX13-600 mmol), HOBt ( has been added. The reaction was kept at -5 °C for 1 hour and then EtOAc (10 mL) and brine (10 mL). The layers were separated and the aqueous layer EtOAc Extracted with (. Organic layers combined and Na2804 dried on. Removed by solid filtration and volatiles removed under reduced pressure. The crude product is compound CX13-600 (98 mg, 1H-NiviR (, 6.57 - 6.73 (m, 2 Synthesis of CX13-601 Scheme 11: Preparing the CX13-601 l'gf'v "J/x _ ..

L x \_ .- l .-Ibn › one-. /xßh ,_ _M rw anti: 0104710 C601-2 CX13-601 Synthesis of C601-1 methylpentanoate ( an addition to the mixture Add DIEA ( at room temperature via funnel. Reaction 16 hours) was kept at room temperature throughout, and then EtOAc (10 mL) and brine (10 mL) was diluted with. The layers were separated and the aqueous layer was extracted with EtOAc ( has been made. The organic layers were combined and dried over Na2804. Thick removed by filtration and volatiles under reduced pressure silica gel to give 601-1 (660 g, 87% yield) as a white solid, which was removed chromatography (PEzEA = *. Synthesis of 0601 -2 stirred under H2 atmosphere throughout. The mixture is filtered and in the next step C m/z.' used without purification 4752(M+HY.

Synthesis of CX13-601 has been added. The reaction was kept at -5 °C for 1 hour and then EtOAc (15 mL) and brine (15 mL). The layers were separated and the aqueous layer EtOAc Extracted with (. Organic layers combined and Na2804 dried on. Removed by solid filtration and volatiles removed under reduced pressure. The crude product contains the compound CX13-601 (70 mg, 1H-NiviR (, 6.89 (d, Synthesis of compound CX13-603 Scheme 12: Preparing the CX13-603 L' i-:aH ri "î/?Ü ' "n , , ..Ph CX13-803 C601-2 mmol), HOBt (69 mg, DIEA ( was added to the mixture. The reaction was maintained at -5 °C for 1 hour and then EtOAc (10 mL) and brine (10 mL). The layers were separated and the aqueous layer EtOAc Extracted with (. Organic layers combined and Na2804 dried on. Removed by solid filtration and volatiles removed under reduced pressure. The crude product is compound CX13-603, (60 mg, Synthesis of compound CX13-605 Scheme 13: Preparing the CX13-605 Is_`~]_`~_/J\Ni/ V _\ 1 > 4 k_ "1 1 f ' "x/'N ..

C104-1Ü .;i fr-s/ luk" TV*** -N 'ii/îl' 3 f/ S +\iî'î - "1 I' g__'i __`-1 "Hi I "J :SH Â \_ `J ,far/M `J V `.1`,.f-`,1 _ L .3 Y" 3 cxiuos '_ 5 Synthesis of C605-2 HBTU (245 mg, 0.77 mmol) into the mixture slowly through an addition funnel. DIEA ( is added. The reaction was kept at room temperature for 16 hours and It was then diluted with EtOAc (10 mL) and brine (10 mL). The layers are separated and the aqueous layer was extracted with EtOAc (. Organic layers combined and dried over Na2804.

Removed by solid filtration and volatiles under reduced pressure removed to give C605-2 (310 g, 89% yield) as a white solid. purified by chromatography. MS (ESI) m/z: *.

Synthesis of C605-3 has been added. The solution was stirred at room temperature for 2 hours. Mixture concentrated and acidified to pH = 5 and yield 0605-3 (150 mg, Preparing the CX13-605 mmol), HOBt ( has been added. The reaction was maintained at -5 °C for 1 hour and EtOAc (10 mL) and brine diluted with water (10 mL). The layers were separated and the aqueous layer EtOAc (with is extracted. Organic layers combined and on N32804 has dried. Removed by solid filtration and volatiles at reduced pressure purified by chromatography. 1H), +.

Synthesis of compound CX13-606 Scheme 14: Preparing the CX13-606 -5 __f-C.. _. &log-R. -.r'i'i'jiH R-x-I Ülîf &cahil-_- L :';" _ HY» -/L`|l" mu_ o . i: 1 ..

-FA i I )-.-/ -~ ~. ;1 -' ii :'; Lar- HC.

ZiCM F A.). C'33î 92./ \`›>.-i'^'=---^*v '~""**_/ “/ Tiiriii s -eru It~ U ü q` .Lux/n - -Nu-'“ÜH . Fi r ~r\ J r 9-.. ` " ceoeia cxusos Synthesis of 0606-4 has been added. The reaction mixture was kept at 0 °C for 2 hours and the volatiles turned white. removed below. MS (ESI) m/z: *. Synthesis of 0606-5 Compound C-2-(tert- mmol), HOBt ( at room temperature).

The reaction was kept at room temperature for 16 hours and then EtOAc (40 mL) and diluted with brine (10 mL). The layers were separated and the aqueous layer was mixed with EtOAc. is extracted. Organic layers combined and on N32804 has dried. Removed by solid filtration and volatiles at reduced pressure removed below to give C606-5 (1.2 9, 89%) as a white solid. purified by chromatography. MS (ESI) m/z: *.

Synthesis of C606-6 Compound C 0 added at °C. The reaction mixture was kept at 0 °C for 2 hours and the volatiles were removed below. MS (ESI) m/z.' *.

Synthesis of C606-7 C133-3 (360) to its mixture in Compound C added at room temperature. The reaction was kept at room temperature for 16 hours. and then diluted with EtOAc (30 mL) and brine (10 mL). The layers are separated and the aqueous layer was extracted with EtOAc (. Organic layers combined and dried over N82804. removed by solid filtration and The volatiles were removed under reduced pressure and produced 608-7ly as a white solid. Synthesis of 0606-8 LiOH.H2O (101 mg, 2.4 mmol) was added at room temperature. My wife for 16 hours mixed at room temperature. Then the mixture was acidified up to pH=5, Extracted with EtOAc (, dried over Na2804. Solid filtration volatiles and 606-8 (440 mg, 92%) as a white solid. yield) was removed under reduced pressure. lVlS (ESI) m/z: *.

Synthesis of CX13-606l To its mixture in Compound C, C103-4 (107 mg, added at the temperature. The reaction was kept at room temperature for 16 hours and It was then diluted with EtOAc (30 mL) and brine (10 mL). The layers are separated and the aqueous layer was extracted with EtOAc (. Organic layers combined and dried over Na2804. removed by solid filtration and The volatiles were removed under reduced pressure and CX13- Purified by chromatography to give 606 well (100 mg, 38%). 1H-NMR (, 7.15 (dd, J = 20.9, Synthesis of compound CX13-608 Scheme 15: Preparing the CX13-608 nar.. en: ~ i :) . H :ii HBTU ( at room temperature has been added. The reaction was kept at room temperature for 16 hours. Later on It was diluted with EtOAc (30 mL) and brine (10 mL). The layers are separated and the aqueous layer Extracted with EtOAC (. Organic layers combined and Na2804 dried on. Removed by solid filtration and volatiles CX1 3-6081 (100 mg, 100 mg) was removed under reduced pressure and as a white solid. EXAMPLE 15Synthesis of Compound CX13-705 Scheme 16: Preparing the CX13-705 1_ l" x/A l'i L:"_›\ !3 "V Wii7 ,J, nm " Etc l: `C2 7 5” MI' C705-1 cms-2 Synthesis of C705-2 The mixture in C 5-methylthiazol-2- carboxylic acid (, HOBt (131 mg, added at room temperature. Reaction 12 hours kept at room temperature throughout. Then EtOAc (20 mL) and brine (10 mL) diluted with. The layers were separated and the aqueous layer was extracted with EtOAc ( has been made. The organic layers were combined and dried over Na2804. Thick removed by filtration and volatiles under reduced pressure removed to give 705-2 (300 mg, 88%) as a white solid. purified by chromatography. LC-MS: m/z +.

Synthesis of C705-3 LiOH.H2O (68mg, 1.62mmol) was added at RT. My wife is in the room for 2 hours. mixed at temp. Then the mixture was acidified to pH=5, EA (20 mL) It was extracted with N82804 and dried over N82804. through solid filtration removed and the volatiles yielded 705-3 (265 mg, 78% yield) as a white solid. removed under reduced pressure for MS (ESI) m/z.' *.

Preparing the CX13-705 mmol), HBTU ( room added at the temperature. The reaction was kept at room temperature for 4 hours. More It was then diluted with EtOAc (20 mL) and brine (10 mL). Layers separated and watery layer was extracted with EtOAc (. The organic layers were combined and It was dried over Na2804. Removed by solid filtration and volatiles Extract CX13-705 (90mg, 90mg) as a white solid which was removed under reduced pressure. The proteasome-activity inhibitory property of the compounds Chymotrypsin-like (CT-L) peptidylglutamyl of 208 proteasomes in enzymatic assays peptide hydrolyzing activity (PGPH) and trypsin-like (T-L) activities. succiniI-Leu-Leu-VaI-Tyr-AMC (10) as substrates with proteasomes, respectively. pmoI/L), Z-Leu-Leu-GIu-AMC (10 µmoI/L) and Boo-Leu-Arg-Arg-AMC (50 µmoI/L) determined using. Stock of 208 proteasome inhibitors described herein solutions were prepared in dimethyl sulfoxide (DMSO) and the final The DMSO concentration was 1%. The reaction is at room temperature for one hour. has been carried out. A plate-based spectrofluorometer of proteasome activity substrates based on the detection of the fluorophore 7-Amino-4-methylcoumarin (AMC) after cleavage with has been measured. IC50 208 is a test in which the catalytic activity of the proteasome is inhibited by 50%. It is a quantitative measure of the concentration of the inhibitor. In certain applications The proteasome inhibitory potential of the compounds is described herein in Table 1.

Water solubility of compounds 1-5 mg of a compound was weighed in a bottle to determine the solubility of the compounds. one ml of aqueous buffer (50 mM citrate at pH 4.0) was added. Bottle at 25 °C for 24 hours It was shaken and then centrifuged at 10000 rpm for 10 minutes.

The supernatant was filtered through a 0.45 µm membrane filter and the compound analyzed for its concentration. In certain applications, the compounds in water The resolution is described here in Table Province.

Table 1. Proteasome activity inhibitor potential (IC50) and water solubility Compound Proteasome inhibition I050 (nM) at pH 4.0 (50 mM) Structure ID number 203 Basic structure 208 proteasome Sequence?" .. in water Immunoproteasome solubility CT-L PGPH T-L CT-L PGPH T-L <9 0 %Hvi o /` Apr' : stv rRi 1 0 H 0 o WNÃÄQJLN& (R) CX13705 >20000 >20000 >20000 15778 CX13709 >20000 >20000 >20000 >20000 Collected data from mice administered a compound described herein at a tolerated dose Inhibition of proteasome activity in snow samples.

BaIb/c mice either a vehicle (20% (w/v) in 10 mM sodium citrate at pH 3.5) hydroxypropyl-p-cyclodextrin) or a compound disclosed herein intravenously at 10 mg/kg a tolerated by injection or oral feeding at 30 mg/kg dose was applied. One hour after dosing, whole blood samples were were collected into tubes containing sodium heparin via xg is centrifuged. The resulting pellet is cold phosphate buffered saline (PBS) It has been used and washed three times. Each time the pellet is reconstituted in 1 ml of cold PBS*. suspended and at 6000 x g for 10 minutes at 4 °C. centrifuged. Cells pelleted after the last wash 100 pL for 1 hour fragmented by addition of lysis buffer (PBS containing 5 mM EDTA, pH 7.4) and then centrifuged at 6000 xg for 10 minutes at 4°C. supernatant transferred to a new tube and the cell pellet discarded. Concentration of blood Iizatin Measured by the BCA method. 10 pg of protein was used for CT-L measurement and pg protein was used for the measurement of PGPH and T-L activities. SukkinI-Leu-Leu- VaI-Tyr-AMC (25 µJmcI/L), Z -Leu-Leu-GIu-AMC (10 µmoI/L) or Boc-Leu-Arg-Arg- AMC (10 pmol/L) is then used as substrate for CT-L, PGPH or T-L, respectively. has been added. The mixture was incubated at 37 °C for 60 minutes. Free AMC to measure the CT-L, PGPH, or T-L activities of the fluorescent 208 proteasome. Measured using a 3601460 nm filter set calibrated on a fluorometer. proteasome percent inhibition of activity from mice administered a compound described herein. comparison of the collected samples with the samples collected from the mice to which the vehicle was administered. calculated by means of Percent inhibition of proteasome activities is shown in FIG. It is explained in 1. Compounds tested as shown in FIG. inhibited CT-L activity in samples. Especially by oral feeding timely, both CT-L and T-L activity of the proteasome in blood samples it inhibits.

In mice bearing human tumor xenografts, anti-tumor compounds of the compounds described herein Female mice (NIH lll HO naked mice purchased from Charles River Laboratories (approximately 20 9, 5-6 weeks old) were kept in cages throughout the experimental period. 100 µL ATCC® (HTB-38T'V') 3x106 human colon cancer cells in phosphate buffer saline (HT-29) or ATCC® (CRL38T'V') 1x107 human B cell lymphoma cells (RL) was injected subcutaneously into the right flank of mice. Mean tumor size animals in such a way that all groups have an equal distribution of tumor size. randomly allocated to different groups (7-10 mice in each group). A vehicle for animals (10%) (by weight/volume) hydroxypropyl-p-cyclodextrin, 10 mM citrate, pH 4.0) or in vehicle by intravenous injection of a prepared solution containing CX13-103 has been applied. Treat animals three times a week on days 1, 2, and 5 for 4-5 weeks. has been applied. Tumor sizes were measured with caliper 2 or 3 every week.

Tumor volume was calculated using the formula: length x widthz/Z. Statistical analysis was obtained using 2-way ANOVA. FIGURE 2A and FIGURE ZBlde As shown, administration of CX13-103 significantly suppressed tumor growth.

(%JaunnxeuewxeÂues HT-29 human colorectal adenocarcinoma xenograft model 2800' -i- Vehicle g 2100-i, g 1400-/1/-541 E 700- :,ß 0 10 20 30 Days after the start of treatment FIGURE 2A RL human Ienfoma xenograft model E 1500- 2 1000 0 10 20 30 Days after the start of treatment

Claims (1)

REQUESTS 1. A compound having the structure shown below and its pharmaceutically acceptable salt or solvate CX13-107. (313-603. A pharmaceutical composition comprising a compound according to claim 1 and a pharmaceutically acceptable carrier. A compound according to claim 1 which specifically inhibits the catalytic activity of 208 proteosamine and inhibits a proteasome-associated disease. or for use in a method of treating the condition. The compound for use according to claim 3, wherein the CT-L activity and T-L activity of the 208 proteasome are simultaneously inhibited. The compound for use according to claim 4, wherein the compound The compound for use according to claim 1, wherein the compound is administered subcutaneously, intravenously, intramuscularly, intraarterially, intrathecally, intracapsularly, intraorbitally, intracardially, intradermally, intraperitoneally, transtracheally. , subcuticularly, intraarticularly, subcapsularly, subarachnoidally, intraspinally, It is administered intraasternally or by infusion. The compound for use according to claim 4, wherein the compound is administered via a non-parenteral route. The compound for use according to claim 7, wherein the compound is administered orally, enterally, buccally, nasally, intranasally, transmucosally, epidermally, transdermally, dermally, ophthalmically, pulmonaryly, sublingually, rectally. It is applied vaginally or topically. The use of a compound according to claim 1 in the manufacture of a medicament for treating a disease or condition associated with 208 proteosome. The compound for use of any of claims 3-8, or the use according to claim 9, wherein the 208 proteasome-related disease or condition is cancer, neurotoxic/degenerative disease, Alzheimer's disease, ischemic condition, inflammation, immune-related disease, HIV infection, organ graft rejection, septic shock, inhibition of antigen presentation, decreased viral gene expression, parasitic infection, acidosis-related condition, macular degeneration, pulmonary condition, muscle atrophy, fibrotic disease, and bone and hair growth diseases.